Korean J Nephrol.  2003 May;22(3):261-272.

Effect of Salicylate on the Monocyte Chemoattractant Protein-1 Expression and Intracellular Reactive Oxygen Species Formation in Human Mesangial Cells

• Chung SJ
• Kim CS
• Chang JW
• Kim SB
• Lee SK
• Park JS

Abstract

BACKGROUND
Monocyte chemoattractant protein-1 (MCP-1) and reactive oxygen species (ROS) play an important role during glomerular inflammation. We investigated the effect of aspirin metabolite, salicylate on the pro-inflammatory cytokine-induced MCP-1 expression and lysophosphatidylcholine -induced intracellular ROS formation in human mesangial cells. METHODS: Cells were pretreated with salicylate, and then stimulated with tumor necrosis factor-alpha (TNF-alpha) or interleukin-1beta (IL-1beta). The expression of MCP-1 mRNA and MCP-1 protein were measured by Northern blot analysis and enzyme-linked immunosorbent assay, respectively. Nuclear factor-kappa B (NF-kappaB) activity was measured by electrophoretic mobility shift assay. Degradation of Ikappa B-alpha was assessed by Western blot analysis. Intracellular ROS production was monitored by flow cytometry using 2'7'-dichlorofluorescin diacetate. RESULTS: Salicylate inhibited the TNF-alpha- or IL-1beta- induced MCP-1 mRNA expression in a dose dependent manner (1-20 mM) and also suppressed the MCP-1 protein expression. Its effect was not attributable to de novo synthesis of intermediary proteins. Salicylate inhibited the TNF-alpha- or IL-1beta-induced NF-kappa B binding activity and also suppressed the TNF-alpha-induced Ikappa B-alpha degradation. Low concentration of salicylate (0.01-1 mM) suppressed the lysophosphatidylcholine-induced ROS formation. CONCLUSION: Milimolar concentration of salicylate inhibited the MCP-1 expression at least in part, via suppression of NF-kappaB by reducing the degradation of Ikappa B-alpha. On the other hand, lower concentration of salicylate could suppress the lysophosphatidylcholine-induced intracellular ROS formation.

Keyword

Mesangial cell; Salicylate; Monocyte chemoattractant protein-1; Nuclear factor-kappaB; Reactive oxygen species

MeSH Terms

Aspirin
Blotting, Northern
Blotting, Western
Chemokine CCL2*
Electrophoretic Mobility Shift Assay
Enzyme-Linked Immunosorbent Assay
Flow Cytometry
Hand
Humans*
Inflammation
Interleukin-1beta
Lysophosphatidylcholines
Mesangial Cells*
Monocytes*
NF-kappa B
Reactive Oxygen Species*
RNA, Messenger
Tumor Necrosis Factor-alpha
Aspirin
Chemokine CCL2
Interleukin-1beta
Lysophosphatidylcholines
NF-kappa B
RNA, Messenger
Reactive Oxygen Species
Tumor Necrosis Factor-alpha